Propofol Differentially Modulates Neurodevelopmental Processes in Region-Specific Human Brain Organoids

Background: Propofol, a commonly used anesthetic, has controversial effects on fetal neurodevelopment. This study explores the hypothesis that propofol exerts region- and time-specific effects on distinct brain regions, with implications for clinical safety during pregnancy. Methods: Human dorsal forebrain (hCS) and ventral forebrain (hSS) organoids were exposed to 20 µM propofol for 6 hours on day 11 (D11). Morphological changes, including diameter and surface area, were analyzed at D11, D18, and D25. Immunofluorescence and PCR analyses were conducted at early (D18/25), middle (D40), and late stages (D80) post-intervention. Electrophysiological analyses were performed from D40 to D70, and bulk RNA-seq analyses were conducted on organoids at D18 to assess transcriptional changes. Results: Propofol exhibited region-specific effects on hCS and hSS organoids. In hCS organoids, excitatory neuron markers were consistently reduced at all stages, while in hSS organoids, inhibitory neuron markers were transiently affected at early stages but recovered over time. Electrophysiological analysis revealed increased action potential incidence in hCS organoids, indicating enhanced neuronal excitability despite decreased excitatory neuron markers. Transcriptomic analysis demonstrated metabolic reprogramming, including improved oxygen supply and a shift toward efficient aerobic pathways, alongside modulation of developmental signaling pathways. Conclusions: Propofol exerts region- and time-specific effects on neural development, involving both neuronal maturation and metabolic adaptation. These findings emphasize the need for careful evaluation of anesthetic safety in pregnant women and its potential long-term neurodevelopmental consequences. Overall design: Human induced pluripotent stem cells (hiPSCs) were differentiated into ventral forebrain organoids and dorsal forebrain organoids, and subjected to propofol exposure on day 11 (D11). Organoids without drug treatment were used as controls. RNA-seq was performed to analyze gene expression profiles, with all samples collected on day 18 (D18) in triplicate.